Hierarchical microspheres with macropores fabricated from chitin as 3D cell culture
Hierarchical microspheres with macropores fabricated from chitin as 3D cell culture
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由甲壳素制成的具有大孔的分层微球作为 3D 细胞培养物
DOI：
10.1039/c9tb01046g
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发表时间：
2019
影响因子：
7
通讯作者：
Zhang Lina
中科院分区：
文献类型：
--
作者：
Su Xiaojuan;Tan Mengtian;Duan Bo;Cai Jie;Jiang Wei;Zhang Lina
3D细胞培养是大规模细胞制造和基于结构的组织工程的迫切需求;然而，由于缺乏用于各种类型细胞的高度生物相容性培养环境，它们的制备仍然具有挑战性。蟹、虾来源的甲壳素具有优良的生物相容性已被证明。本文首次以壳聚糖微球为牺牲模板，基于其选择性溶解性，设计并制备了具有开放可调大孔的分级甲壳素微球。将不同尺寸的壳聚糖微球包埋在由NaOH/尿素水溶液制备的甲壳素微球中，然后用弱酸去除，形成平均孔径为63 μm、94 μm和135 μm的大孔。重要的是，由于纳米纤维网络的存在，具有大孔结构的平均直径为240 μm的甲壳素微球表现出良好的强度。由于甲壳素本身的生物活性，甲壳素纳米纤维在增强PCMS的力学性能和促进细胞的粘附、生长和增殖方面发挥了重要作用。此外，甲壳素大孔微球可以支持多种哺乳动物细胞的生长，如猴成纤维细胞、人肝细胞甚至人干细胞。在该微球中培养的干细胞保持了其三维结构和多谱系分化能力。本研究提供了一种新的“自下而上”的方法，从甲壳素溶液中创建大孔微球，这可用于大规模的人类干细胞三维培养，也是一个有前途的候选人，为制造优良的支架。
3D cell culture is an urgent requirement for large-scale cell manufacture and structure-based tissue engineering; however, their preparation is still challenging due to the lack of a highly biocompatible culture environment for various types of cells. The excellent biocompatibility of chitin derived from crabs and shrimps has been proved before. Herein, for the first time, we designed and prepared hierarchical chitin microspheres with open tunable macropores using chitosan microspheres as a sacrificial template, based on their selective solubility. In our findings, the chitosan microspheres with various sizes were embedded in the chitin microsphere fabricated from NaOH/urea aqueous solution, and then were removed using a weak acid to form macropores with mean pore sizes of 63 μm, 94 μm and 135 μm. Importantly, the chitin microspheres having an average diameter of 240 μm with the macroporous structure exhibited good strength, as a result of the existence of nanofibrous networks. The chitin nanofibers played important roles in both enhancing the mechanical properties of PCMS and improving the adhesion, growth, and proliferation of cells due to the inherent bioactivity of chitin. Moreover, the chitin macroporous microspheres could support the growth of various mammalian cells, such as monkey fibroblasts, human hepatocytes and even human stem cells. The stem cells cultured in this microsphere maintained their 3D structure and multi-lineage differentiation capacity. This study provides a new “bottom-up” approach to create macroporous microspheres from a chitin solution, which could be used for large-scale human stem cell 3D culture and also as a promising candidate for the fabrication of excellent scaffolds.